对暴露于地西他滨的细胞进行发夹 - 亚硫酸氢盐测序记录了 DNA 去甲基化的过程。
Hairpin-bisulfite sequencing of cells exposed to decitabine documents the process of DNA demethylation.
机构信息
Department of Pathology, University of Otago, Dunedin, New Zealand.
Department of Biochemistry, University of Otago, Dunedin, New Zealand.
出版信息
Epigenetics. 2021 Nov;16(11):1251-1259. doi: 10.1080/15592294.2020.1861169. Epub 2020 Dec 28.
Abstract
Although the mechanism of DNA demethylating drugs has been understood for many years, the direct effect of these drugs on methylation of the complementary strands of DNA has not been formally demonstrated. By using hairpin-bisulphite sequencing, we describe the kinetics and pattern of DNA methylation following treatment of cells by the DNA methyltransferase 1 (DNMT1) inhibitor, decitabine. As expected, we demonstrate complete loss of methylation on the daughter strand following S-phase in selected densely methylated genes in synchronized Jurkat cells. Thereafter, cells showed a heterogeneous pattern of methylation reflecting replication of the unmethylated strand and restoration of methylation.
摘要
尽管 DNA 去甲基化药物的作用机制已经被人们了解多年，但这些药物对 DNA 互补链甲基化的直接作用尚未得到正式证实。通过使用发夹 - 亚硫酸氢盐测序，我们描述了在细胞用 DNA 甲基转移酶 1（DNMT1）抑制剂地西他滨处理后 DNA 甲基化的动力学和模式。正如预期的那样，我们在同步化的 Jurkat 细胞中选择的高度甲基化基因中，在 S 期后观察到子链上的甲基化完全丢失。此后，细胞显示出异质的甲基化模式，反映了未甲基化链的复制和甲基化的恢复。
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